Seedling and adult‐plant stage resistance of a world collection of barley genotypes to stripe rust

Barley stripe rust caused by Puccinia striiformis f.sp. hordei (PSH) is one of the major diseases in barley production regions worldwide. A total of 336 barley genotypes with diverse genetic backgrounds targeted for low‐input barley production were tested for seedling and adult‐plant stage resistance against six PSH races (0S0, 0S0‐1, 1S0, 4S0, 5S0 and 7S0) originated from India. The seedling resistance was evaluated by inoculating the barley genotypes with six races separately under controlled conditions in Shimla, India. The same barley genotypes were evaluated for adult‐plant stage resistance in the Agricultural Research Station (ARS) of Rajasthan Agriculture University, Durgapura, Rajasthan, India. Out of the 336 barley genotypes tested for seedling resistance, 119 (35.4%), 101 (30.1%), 87 (25.9%), 100 (29.8%), 91 (27.1%) and 70 (20.8%) genotypes were resistant to races 0S0, 0S0‐1, 1S0, 4S0, 5S0 and 7S0, respectively. In the field, 102 (30.3%) genotypes showed the resistance response of which 18 (5.3%) genotypes were highly resistant to PSH. Barley genotypes AM‐14, AM‐177, AM‐37, AM‐120, AM‐300, AM‐36, AM‐103, AM‐189, AM‐291, AM‐275 and AM‐274 showed resistance response to all six races at seedling and adult‐plant stages. Seedling resistance reported in the current study is effective against the newly emerged race 7S0 and previously reported five races in India. Therefore, resistant barley genotypes identified in the current study provided effective protection against all six races at seedling and adult‐plant stages. The stripe rust resistance identified in the current studies may be potential donors of stripe rust resistance to barley breeding programmes in India and elsewhere.     

不同疗程全身糖皮质激素对AECOPD患者的疗效与安全性Meta分析

目的通过系统评价比较不同疗程全身糖皮质激素治疗对AECOPD患者的有效性与安全性。方法计算机检索PubMed,EMBASE数据库,Cochrane图书馆中关于对AECOPD患者全身使用糖皮质激素的所有文献(使用随机对照实验)。检索时间从建库至2016年1月,并由2名评价者独立筛选文献、提取资料并评估纳入研究偏倚风险后,采用RevMan 5.0软件进行Meta分析。激素组按照疗程分为短期组(≤3天)和长期组(3天),分别在两组中按照起始剂量80mg泼尼松为界限分为高剂量组(≥80mg/d)和低剂量组(80mg/d)两个亚组,并进行分析。结果纳入文献11篇,涉及1142例患者。其中10篇文献涉及肺功能FEV1,在短期组中使用SCS后病人FEV1明显提高,Meta分析结果显示[MD=0.13,95%CI(0.08,019)],在短期组的中高剂量组为[MD=0.09,95%CI(0.03,0.16)],低剂量组为[MD=0.23,95%CI(0.12,0.33)],差异均有统计学意义。长期组与对照组比较,FEV1差异无统计学意义[MD=0.06,95%CI(-0.01,012)];在长期组的中低剂量组与对照组比较,差异有统计学意义[MD=0.14,95%CI(0.05,0.22)],高剂量组无统计学意义[MD=-0.06,95%CI(-0.16,0.04)]。激素组与对照组不良反应患者中,发生高血糖的Meta分析结果显示[OR=3.94,95%CI(2.15,7.24)],高剂量组[OR=3.50,95%CI(1.73,7.05)],低剂量组[OR=5.48,95%CI(1.58,18.96)],高低剂量激素组发生糖尿病的风险均较对照组增加,差异均有统计学意义。激素组与对照组及不同剂量亚组中二重感染、高血压、胃肠道出血等不良反应比较,差异无统计学意义。结论AECOPD患者行全身糖皮质激素治疗以低剂量短期疗法的效果较好,安全性更高。     

Solution structure of lysine‐free (K0) ubiquitin

Lysine‐free ubiquitin (K0‐Ub) is commonly used to study the ubiquitin‐signaling pathway, where it is assumed to have the same structure and function as wild‐type ubiquitin (wt‐Ub). However, the K0‐Ub ¹⁵N heteronuclear single quantum correlation NMR spectrum differs significantly from wt‐Ub and the melting temperature is depressed by 19°C, raising the question of the structural integrity and equivalence to wt‐Ub. The three‐dimensional structure of K0‐Ub was determined by solution NMR, using chemical shift and residual dipolar coupling data. K0‐Ub adopts the same backbone structure as wt‐Ub, and all significant chemical shifts can be related to interactions impacted by the K to R mutations.     

The effects of light intensity and algae-induced turbidity on feeding behaviour of larval striped trumpeter

Striped trumpeter larvae reared in algal cell‐induced turbid water (greenwater) fed equally well in clearwater in a light intensity range of 1–10 μmol s^‐1 m^‐2, when evaluated in terms of both the proportion of larvae feeding and larval feeding intensity. An ontogenetic improvement in photopic visual sensitivity of larvae was indicated by improved feeding at 0·1 μmol s^‐1 m^‐2, from 26±5% of larvae feeding and 0·027±0·005 rotifers consumed per feeding larva min^‐1 on day 8, to 96±2% and 0·221±0·007 rotifers consumed larva^‐1 min^‐1 on day 23 post‐hatching. Algal cell‐induced turbidity was shown to reduce incident irradiance with depth, indicated by increasing coefficients of attenuation (1·4–33·1) with increasing cell densities (0–2×10⁶ cells ml^‐1), though light intensities in the feeding experiment test chambers, at the algal cell densities tested, were within the optimal range for feeding (1–10 μmol s^‐1 m^‐2). Algae‐induced turbidity had different effects on larval feeding response dependent upon the previous visual environment of the larvae. Young larvae (day 9 post‐hatching) reared in clearwater showed decreased feeding capabilities with increasing turbidity, from 98±1% feeding and 0·153±0·022 rotifers consumed larva^‐1 min^‐1 in clearwater to 61±10% feeding and 0·042±0·004 rotifers consumed larva^‐1 min^‐1 at 56 NTU, while older clearwater reared larvae fed well at all turbidities tested. Likewise, greenwater reared larvae had increased feeding capabilities in the highest algal cell densities tested (32 and 66 NTU) compared with those in low algal cell density (6 NTU), and clearwater (0·7 NTU) to which they were naïve.     

Effects of extracellular matrix proteins in chondrocyte‐derived matrices on chondrocyte functions

Loss of cartilaginous phenotype during in vitro expansion culture of chondrocytes is a major barrier to the application of chondrocytes for tissue engineering. In previous study, we showed that dedifferentiation of chondrocytes during the passage culture was delayed by matrices formed by primary chondrocytes (P0‐ECM). In this study, we investigated bovine chondrocyte functions when being cultured on isolated extracellular matrix (ECM) protein‐coated substrata and P0‐ECM. Low chondrocyte attachment was observed on aggrecan‐coated substratum and P0‐ECM. Cell proliferation on aggrecan‐ and type II collagen/aggrecan‐coated substrata and P0‐ECM was lower than that on the other ECM protein (type I collagen and type II collagen)‐coated substrata. When chondrocytes were subcultured on aggrecan‐coated substratum, decline of cartilaginous gene expression was delayed, which was similar to the cells subcultured on P0‐ECM. These results indicate that aggrecan plays an important role in the regulation of chondrocyte functions and P0‐ECM may be a good experimental control for investigating the role of each ECM protein in cartilage ECM. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1331–1336, 2013     

Inhibition of MCL‐1 by obatoclax sensitizes Sp2/0‐Ag14 hybridoma cells to glutamine deprivation‐induced apoptosis

For several cancer cell types, the lack of an adequate supply of the amino acidl ‐glutamine (Gln) triggers apoptosis, a phenomenon termed Gln addiction. In this report, we examined the role of the anti‐apoptotic proteins of the B‐cell lymphoma 2 (BCL‐2) protein family in the survival of Sp2/0‐Ag14 (Sp2/0) mouse hybridoma cells, a cell line that undergoes apoptosis within minutes of Gln deprivation. Western blot analysis revealed that myeloid cell leukaemia 1 (MCL‐1) was expressed at much higher levels than BCL‐2, B‐cell lymphoma extra‐large and BCL‐2‐like protein 2 making it the prominent pro‐survival BCL‐2 family member in this hybridoma. Gln deprivation triggered a progressive decrease in MCL‐1 protein levels, which coincided with the decrease in Sp2/0 cell survival. Moreover, Sp2/0 cells were much more sensitive to the broad Bcl‐2 homology domain‐3 (BH3) mimetic obatoclax (which targets MCL‐1) than to the more selective drug ABT‐737 (which does not target MCL‐1). Finally, we show that obatoclax sensitizes Sp2/0 cells to apoptosis following Gln starvation. All together, the data presented here reveal that modulation of the pro‐survival protein MCL‐1 is an important step in the sequence of events leading to the initiation of apoptosis in Gln‐starved Sp2/0 cells. Cancer cells require an adequate supply ofl ‐glutamine for their survival. Using a mouse hybridoma cell line that is exquisitely sensitive to glutamine starvation, we show that the levels of the pro‐survival BCL‐2 family protein MCL‐1 decrease upon glutamine starvation in a manner that correlates with the loss of cell viability. Moreover, inhibiting MCL‐1 with the drug obatoclax sensitizes hybridoma cells to glutamine starvation. Thus, in some cancer cells, glutamine starvation triggers the inactivation of pro‐survival proteins. Our data suggest that the combined inhibition of glutamine biosynthesis pathways and BCL‐2 proteins may prove effective against some cancers. Copyright © 2015 John Wiley & Sons, Ltd.     

Physical training prevents oxidative stress in L‐NAME‐induced hypertension rats

The present study investigated the effects of a 6‐week swimming training on blood pressure, nitric oxide (NO) levels and oxidative stress parameters such as protein and lipid oxidation, antioxidant enzyme activity and endogenous non‐enzymatic antioxidant content in kidney and circulating fluids, as well as on serum biochemical parameters (cholesterol, triglycerides, urea and creatinine) from Nω‐nitro‐L‐arginine methyl ester hydrochloride (L‐NAME)‐induced hypertension treated rats. Animals were divided into four groups (n = 10): Control, Exercise, L‐NAME and Exercise L‐NAME. Results showed that exercise prevented a decrease in NO levels in hypertensive rats (P < 0·05). An increase in protein and lipid oxidation observed in the L‐NAME‐treated group was reverted by physical training in serum from the Exercise L‐NAME group (P < 0·05). A decrease in the catalase (CAT) and superoxide dismutase (SOD) activities in the L‐NAME group was observed when compared with normotensive groups (P < 0·05). In kidney, exercise significantly augmented the CAT and SOD activities in the Exercise L‐NAME group when compared with the L‐NAME group (P < 0·05). There was a decrease in the non‐protein thiols (NPSH) levels in the L‐NAME‐treated group when compared with the normotensive groups (P < 0·05). In the Exercise L‐NAME group, there was an increase in NPSH levels when compared with the L‐NAME group (P < 0·05). The elevation in serum cholesterol, triglycerides, urea and creatinine levels observed in the L‐NAME group were reverted to levels close to normal by exercise in the Exercise L‐NAME group (P < 0·05). Exercise training had hypotensive effect, reducing blood pressure in the Exercise L‐NAME group (P < 0·05). These findings suggest that physical training could have a protector effect against oxidative damage and renal injury caused by hypertension. Copyright © 2012 John Wiley & Sons, Ltd.     

A revised model for the control of fatty acid synthesis by master regulator Spo0A in Bacillus subtilis

In starving Bacillus subtilis cells, the accDA operon encoding two subunits of the essential acetyl‐CoA carboxylase (ACC) has been proposed to be tightly regulated by direct binding of the master regulator Spo0A to a cis element (0A box) in the promoter region. When the 0A box is mutated, biofilm formation and sporulation have been reported to be impaired. Here, we present evidence that two 0A boxes, one previously known (0A‐1) and another newly discovered (0A‐2) in the accDA promoter region are positively and negatively regulated by Spo0A～P respectively. Cells with mutated 0A boxes experience slight delays in sporulation, but eventually sporulate with high efficiency. In contrast, cells harboring a single mutated 0A‐2 box are deficient for biofilm formation, while cells harboring either a mutated 0A‐1 box or both mutated 0A boxes form biofilms. We further show that the essential ACC enzyme localizes on or near the cell membrane by directly observing a functional GFP fusion to one of the enzyme's subunits. Collectively, we propose a revised model in which accDA is primarily transcribed by a major σ^A‐RNA polymerase, while Spo0A～P plays an additional role in the fine‐tuning of accDA expression upon starvation to support proper biofilm formation and sporulation.     

Redirection of metabolic flux for high levels of omega‐7 monounsaturated fatty acid accumulation in camelina seeds

Seed oils enriched in omega‐7 monounsaturated fatty acids, including palmitoleic acid (16:1?9) and cis‐vaccenic acid (18:1?11), have nutraceutical and industrial value for polyethylene production and biofuels. Existing oilseed crops accumulate only small amounts (<2%) of these novel fatty acids in their seed oils. We demonstrate a strategy for enhanced production of omega‐7 monounsaturated fatty acids in camelina (Camelina sativa) and soybean (Glycine max) that is dependent on redirection of metabolic flux from the typical ?9 desaturation of stearoyl (18:0)‐acyl carrier protein (ACP) to ?9 desaturation of palmitoyl (16:0)‐acyl carrier protein (ACP) and coenzyme A (CoA). This was achieved by seed‐specific co‐expression of a mutant ?9‐acyl‐ACP and an acyl‐CoA desaturase with high specificity for 16:0‐ACP and CoA substrates, respectively. This strategy was most effective in camelina where seed oils with ~17% omega‐7 monounsaturated fatty acids were obtained. Further increases in omega‐7 fatty acid accumulation to 60–65% of the total fatty acids in camelina seeds were achieved by inclusion of seed‐specific suppression of 3‐keto‐acyl‐ACP synthase II and the FatB 16:0‐ACP thioesterase genes to increase substrate pool sizes of 16:0‐ACP for the ?9‐acyl‐ACP desaturase and by blocking C18 fatty acid elongation. Seeds from these lines also had total saturated fatty acids reduced to ~5% of the seed oil versus ~12% in seeds of nontransformed plants. Consistent with accumulation of triacylglycerol species with shorter fatty acid chain lengths and increased monounsaturation, seed oils from engineered lines had marked shifts in thermotropic properties that may be of value for biofuel applications.     

VHS domains of ESCRT‐0 cooperate in high‐avidity binding to polyubiquitinated cargo

VHS (Vps27, Hrs, and STAM) domains occur in ESCRT‐0 subunits Hrs and STAM, GGA adapters, and other trafficking proteins. The structure of the STAM VHS domain–ubiquitin complex was solved at 2.6 Å resolution, revealing that determinants for ubiquitin recognition are conserved in nearly all VHS domains. VHS domains from all classes of VHS‐domain containing proteins in yeast and humans, including both subunits of ESCRT‐0, bound ubiquitin in vitro. ESCRTs have been implicated in the sorting of Lys63‐linked polyubiquitinated cargo. Intact human ESCRT‐0 binds Lys63‐linked tetraubiquitin 50‐fold more tightly than monoubiquitin, though only 2‐fold more tightly than Lys48‐linked tetraubiquitin. The gain in affinity is attributed to the cooperation of flexibly connected VHS and UIM motifs of ESCRT‐0 in avid binding to the polyubiquitin chain. Mutational analysis of all the five ubiquitin‐binding sites in yeast ESCRT‐0 shows that cooperation between them is required for the sorting of the Lys63‐linked polyubiquitinated cargo Cps1 to the vacuole.     

Specific and spatial labeling of P0‐Cre versus Wnt1‐Cre in cranial neural crest in early mouse embryos

P0‐Cre and Wnt1‐Cre mouse lines have been widely used in combination with loxP‐flanked mice to label and genetically modify neural crest (NC) cells and their derivatives. Wnt1‐Cre has been regarded as the gold standard and there have been concerns about the specificity of P0‐Cre because it is not clear about the timing and spatial distribution of the P0‐Cre transgene in labeling NC cells at early embryonic stages. We re‐visited P0‐Cre and Wnt1‐Cre models in the labeling of NC cells in early mouse embryos with a focus on cranial NC. We found that R26‐lacZ Cre reporter responded to Cre activity more reliably than CAAG‐lacZ Cre reporter during early embryogenesis. Cre immunosignals in P0‐Cre and reporter (lacZ and RFP ) activity in P0‐Cre/R26‐lacZ and P0‐Cre/R26‐RFP embryos was detected in the cranial NC and notochord regions in E8.0–9.5 (4–19 somites) embryos. P0‐Cre transgene expression was observed in migrating NC cells and was more extensive in the forebrain and hindbrain but not apparent in the midbrain. Differences in the Cre distribution patterns of P0‐Cre and Wnt1‐Cre were profound in the midbrain and hindbrain regions, that is, extensive in the midbrain of Wnt1‐Cre and in the hindbrain of P0‐Cre embryos. The difference between P0‐Cre and Wnt1‐Cre in labeling cranial NC may provide a better explanation of the differential distributions of their NC derivatives and of the phenotypes caused by Cre‐driven genetic modifications.     

Priority effects among young‐of‐the‐year fish: reduced growth of bluegill sunfish (Lepomis macrochirus) caused by yellow perch (Perca flavescens)?

1. When available, Daphnia spp. are often preferred by age‐0 yellow perch and bluegill sunfish because of energetic profitability. We hypothesised that predation by age‐0 yellow perch could lead to a midsummer decline (MSD) of Daphnia spp. and that priority effects may favour yellow perch because they hatch before bluegill, allowing them to capitalise on Daphnia spp. prior to bluegill emergence. 2. Data were collected from 2004 to 2010 in Pelican Lake, Nebraska, U.S.A. The lake experienced a prolonged MSD in all but 1 year (2005), generally occurring within the first 2 weeks of June except in 2008 and 2010 when it occurred at the end of June. MSD timing is not solely related to seasonal patterns of age‐0 yellow perch consumption. Nevertheless, when Daphnia spp. biomass was low during 2004 and 2006–2010 (<4 mg wet weight L^?1), predation by age‐0 yellow perch seems to have suppressed Daphnia spp. biomass (i.e. <1.0 mg wet weight L^?1). The exception was 2005 when age‐0 yellow perch were absent. 3. Growth of age‐0 bluegill was significantly faster in 2005, when Daphnia spp. were available in greater densities (>4 mg wet weight L^?1) compared with the other years (<0.2 mg wet weight L^?1). 4. We conclude that age‐0 yellow perch are capable of reducing Daphnia biomass prior to the arrival of age‐0 bluegill, ultimately slowing bluegill growth. Thus, priority effects favour age‐0 yellow perch when competing with age‐0 bluegill for Daphnia. However, these effects may be minimised if there is a shorter time between hatching of the two species, higher Daphnia spp. densities or lower age‐0 yellow perch densities.     

Suppression of oxidative phosphorylation confers resistance against bevacizumab in experimental glioma

Although bevacizumab initially shows high response rates in gliomas and other tumours, therapy resistance usually develops later. Because anti‐angiogenic agents are supposed to induce hypoxia, we asked whether rendering glioma cells independent of oxidative phosphorylation modulates their sensitivity against hypoxia and bevacizumab. LNT‐229 glioma cells without functional mitochondria (rho⁰) and control (rho⁺) cells were generated. LNT‐229 rho⁰‐cells displayed reduced expression of oxidative phosphorylation‐related genes and diminished oxygen consumption. Conversely, glycolysis was up‐regulated in these cells, as shown by increased lactate production and stronger expression of glucose transporter‐1 and lactate dehydrogenase‐A. However, hypoxia‐induced cell death in vitro was nearly completely abolished in the LNT‐229 rho⁰‐cells, these cells were more sensitive towards glucose restriction and the treatment with the glycolysis inhibitor 2‐deoxy‐D‐glucose. In an orthotopic mouse xenograft experiment, bevacizumab induced hypoxia as reflected by elevated Hypoxia‐inducible factor 1‐alpha staining in both, rho⁺‐ and rho⁰‐tumours. However, it prolonged survival only in the mice bearing rho⁺‐tumours (74 days vs. 105 days, p  = 0.024 log‐rank test) and had no effect on survival in mice carrying LNT‐229 rho⁰‐tumours (75 days vs. 70 days, p  = 0.52 log‐rank test). Interestingly, inhibition of glycolysis in vivo with 2‐deoxy‐D‐glucose re‐established sensitivity of rho⁰‐tumours against bevacizumab (98 days vs. 80 days, p  = 0.0001). In summary, ablation of oxidative phosphorylation in glioma cells leads to a more glycolytic and hypoxia‐resistant phenotype and is sufficient to induce bevacizumab‐refractory tumours. These results add to increasing evidence that a switch towards glycolysis is one mechanism how tumour cells may evade anti‐angiogenic treatments and suggest anti‐glycolytic strategies as promising approaches to overcome bevacizumab resistance.

     

Staphylococcus aureus‐mediated blood–brain barrier injury: an in vitro human brain microvascular endothelial cell model

Blood–brain barrier (BBB) disruption constitutes a hallmark event during pathogen‐mediated neurological disorders such as bacterial meningitis. As a prevalent opportunistic pathogen, Staphylococcus aureus (SA) is of particular interest in this context, although our fundamental understanding of how SA disrupts the BBB is very limited. This paper employs in vitro infection models to address this. Human brain microvascular endothelial cells (HBMvECs) were infected with formaldehyde‐fixed (multiplicity of infection [MOI] 0–250, 0–48 hr) and live (MOI 0–100, 0–3 hr) SA cultures. Both Fixed‐SA and Live‐SA could adhere to HBMvECs with equal efficacy and cause elevated paracellular permeability. In further studies employing Fixed‐SA, infection of HBMvECs caused dose‐dependent release of cytokines/chemokines (TNF‐α, IL‐6, MCP‐1, IP‐10, and thrombomodulin), reduced expression of interendothelial junction proteins (VE‐Cadherin, claudin‐5, and ZO‐1), and activation of both canonical and non‐canonical NF‐κB pathways. Using N‐acetylcysteine, we determined that these events were coupled to the SA‐mediated induction of reactive oxygen species (ROS) within HBMvECs. Finally, treatment of HBMvECs with Fixed‐ΔSpA (MOI 0–250, 48 hr), a gene deletion mutant of Staphylococcal protein A associated with bacterial infectivity, had relatively similar effects to Newman WT Fixed‐SA. In conclusion, these findings provide insight into how SA infection may activate proinflammatory mechanisms within the brain microvascular endothelium to elicit BBB failure.     

Large gradient high magnetic field affects the association of MACF1 with actin and microtubule cytoskeleton

The intense inhomogeneous magnetic fields acting on the diamagnetic materials naturally present in cells can generate strong magnetic forces. We have developed a superconducting magnet platform with large gradient high magnetic field (LG‐HMF), which can produce three magnetic force fields of ?1360, 0, and 1312 T²/m, and three corresponding apparent gravity levels, namely 0, 1, and 2‐g for diamagnetic materials. In this study, the effects of different magnetic force fields on osteoblast‐like cells (MG‐63 and MC3T3‐E1) viability, microtubule actin crosslinking factor 1 (MACF1) expression and its association with cytoskeleton were investigated. Results showed that cell viability increased to different degrees after exposure to 0 or 1‐g conditions for 24 h, but it decreased by about 30% under 2‐g conditions compared with control conditions. An increase in MACF1 expression at the RNA or protein level was observed in osteoblast‐like cells under the magnetic force field of ?1360 T²/m (0‐g) relative to 1312 T²/m (2‐g). Under control conditions, anti‐MACF1 staining was scattered in the cytoplasm and partially colocalized with actin filaments (AFs) or microtubules (MTs) in the majority of osteoblast‐like cells. Under 0‐g conditions, MACF1 labeling was concentrated at perinuclear region and colocalization was not apparent. The patterns of anti‐MACF1 labeling on MTs varied with MTs' changing under LG‐HMF environment. In conclusion, LG‐HMF affects osteoblast‐like cell viability, MACF1 distribution, expression, and its association with cytoskeleton to some extent. Bioelectromagnetics 30:545–555, 2009. © 2009 Wiley‐Liss, Inc.     

Effects of dietary l-tryptophan and lighting conditions on growth performance of European sea bass (Dicentrarchus labrax) juveniles reared in a recirculating water system

The aim of the present study was to investigate possible stressful effects on European sea bass Dicentrarchus labrax reared under constant darkness (0L‐24D) and to examine the possible anti‐stressful effect of dietary tryptophan (TRP) supplementation. Juvenile European sea bass (initial body weight 4.23 ± 0.032 g) were reared for 10 weeks under 0L‐24D and 12L‐12D and fed either a commercial diet (0.47% TRP) or the same diet supplemented with L‐TRP (2.47% TRP). Results showed that lighting conditions had no significant effect on fish growth, while a depressive effect by the TRP supplementation was obvious. All fish populations reared under 0L‐24D exhibited reduced body protein, lipid and ash content and increased food consumption. Reduced body lipids, food consumption and nutrient utilization were observed in TRP‐supplemented fed fish, along with lower liver lipids. Dietary TRP enrichment significantly lowered liver saturated and monounsaturated acids and increased poly‐ and highly‐unsaturated fatty acids, especially in combination with 0L‐24D. These changes were also considerably reflected in carcass fatty acid composition.     

乳腺导管原位癌患者钼靶摄影表现与ki-67、PR、ER、HER-2表达的相关性

目的探讨乳腺导管原位癌(DCIS)患者钼靶摄影表现与ki-67、PR、ER、HER-2表达的相关性。方法回顾性分析2010年1月~2016年1月入住三明市第二医院普外科经病理检查明确诊断为DCIS的患者82例,收集82例DCIS患者的年龄、身高、体重、是否绝经、病理资料、乳腺钼靶等资料,根据乳腺钼靶摄影有无钙化分为钙化组与非钙化组,并分析两组患者的年龄、体质指数、是否绝经、乳腺钼靶表现与DCIS患者ki-67、PR、ER、HER-2表达的相关性。结果非钙化DCIS患者ki-67阳性率为58.3%(21/36),PR阳性率为52.8%(19/36),ER阳性率为55.6%(20/36),HER-2阳性率为38.7%(14/36),钙化DCIS患者的ki-67、HER-2阳性率较非钙化患者均明显升高。随着年龄的增长,DCIS患者的ki-67阳性表达率降低,HER-2阳性表达率升高;随着BMI的增加,DCIS患者的PR、ER阳性表达率亦逐渐升高;而未绝经的DCIS患者PR、ER阳性表达率较已绝经患者均明显升高。结论乳腺钼靶摄影表现为钙化的DCIS患者ki-67高表达,其HER-2亦高表达,以上特征可在一定程度上指导临床制定个体化诊治方案。     

Hedge Mustard (Sisymbrium officinale): Chemical Diversity of Volatiles and Their Antimicrobial Activity

Volatile compounds of hedge mustard (Sysimbrium officinale) have been investigated for the first time. Forthy‐two compounds were identified after hydrodistillation (without or upon autolysis) after gas chromatography and gas chromatography/mass spectrometry analyses. In addition, after decoction and hydrolysis of O‐glycosides, 18 volatile O‐aglycones were identified. In general, the main volatiles found in hydrodistillates were: isopropyl isothiocyanate (27.6–48.9%), 2‐methylpropanenitrile (0.5–18.8%), (Z)‐hex‐3‐en‐1‐ol (0.5–18.0%), sec‐butyl isothiocyanate (4.9–9.4%), (E)‐hex‐2‐enal (3.5–8.6%), (Z)‐hex‐2‐en‐1‐ol (0.3–8.4%), octanoic (0.5–8.6%) and dodecanoic acid (0–5.0%), 2‐methylbutanenitrile (0–4.6%), dibutyl phthalate (0–4.5%), and ethyl linolenate (0–3.6%). The main volatile O‐aglycones were: 2‐phenylethyl alcohol (21.5%), 6,7‐dehydro‐7,8‐dihydro‐3‐oxo‐α‐ionol (9.3%), eugenol (8.3%), benzyl alcohol (7.0%), ethyl vanillate (5.2%), 6‐(tert‐butyl)‐5‐methylphenol (5.1%), vanillin acetone (4.7%), ethyl 4‐hydroxybenzoate (4.3%), and 2‐hydroxy‐β‐ionone (3.8%). All hydrodistillates exhibited great potential of antibacterial activity against five Gram‐positive bacteria, nine ampicillin‐resistant Gram‐negative bacteria, and four fungi at a concentration of 500 μg/ml using the disc diffusion method.     

Structural basis for endosomal recruitment of ESCRT-I by ESCRT-0 in yeast

The ESCRT‐0 and ESCRT‐I complexes coordinate the clustering of ubiquitinated cargo with intralumenal budding of the endosomal membrane, two essential steps in vacuolar/lysosomal protein sorting from yeast to humans. The 1.85‐Å crystal structure of interacting regions of the yeast ESCRT‐0 and ESCRT‐I complexes reveals that PSDP motifs of the Vps27 ESCRT‐0 subunit bind to a novel electropositive N‐terminal site on the UEV domain of the ESCRT‐I subunit Vps23 centred on Trp16. This novel site is completely different from the C‐terminal part of the human UEV domain that binds to P(S/T)AP motifs of human ESCRT‐0 and HIV‐1 Gag. Disruption of the novel PSDP‐binding site eliminates the interaction in vitro and blocks enrichment of Vps23 in endosome‐related class E compartments in yeast cells. However, this site is non‐essential for sorting of the ESCRT cargo Cps1. Taken together, these results show how a conserved motif/domain pair can evolve to use strikingly different binding modes in different organisms.     

Maternal hypoxia increases the activity of MMPs and decreases the expression of TIMPs in the brain of neonatal rats

A recent study has shown that increased activity of matrix metalloproteinases‐2 and metalloproteinases‐9 (MMP‐2 and MMP‐9) has detrimental effect on the brain after neonatal hypoxia. The present study determined the effect of maternal hypoxia on neuronal survivability and the activity of MMP‐2 and MMP‐9, as well as the expression of tissue inhibitors of metalloproteinase 1 and 2 (TIMP‐1 and TIMP‐2) in the brain of neonatal rats. Pregnant rats were exposed to 10.5% oxygen for 6 days from the gestation day 15 to day 21. Pups were sacrificed at day 0, 4, 7, 14, and 21 after birth. Body weight and brain weight of the pups were measured at each time point. The activity of MMP‐2 and MMP‐9 and the protein abundance of TIMP‐1 and TIMP‐2 were determined by zymography and Western blotting, respectively. The tissue distribution of MMPs was examined by immunofluorescence staining. The neuronal death was detected by Nissl staining. Maternal hypoxia caused significant decreases in body and brain size, increased activity of MMP‐2 at day 0, and increased MMP‐9 at day 0 and 4. The increased activity of the MMPs was accompanied by an overall tendency towards a reduced expression of TIMPs at all ages with the significance observed for TIMPs at day 0, 4, and 7. Immunofluorescence analysis showed an increased expression of MMP‐2, MMP‐9 in the hippocampus at day 0 and 4. Nissl staining revealed significant cell death in the hippocampus at day 0, 4, and 7. Functional tests showed worse neurobehavioral outcomes in the hypoxic animals. © 2009 Wiley Periodicals, Inc. Develop Neurobiol 2010